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. 2020 Nov 3;11:536442. doi: 10.3389/fimmu.2020.536442

Figure 3.

Figure 3

Heterozygous deletion of A20 affects hematopoietic stem and progenitor cell frequency. (A) Outline of experiments utilizing WT (VavCre+ or RosaCreER+) and A20+/- (A20+/-VavCre+ or A20+/-RosaCreER+) mice for NF-κB signaling and HSPC analysis. (B–D) Summary of BM HSPC proportions at 16 weeks after transplantation using VavCre+ (n = 6) or A20+/-VavCre+ BM cells (n = 8). (E) Flow cytometric analysis of RosaCreER+ and A20+/-RosaCreER+ LSK BM treated with 10 μM EdU for 5 h (LSK pooled from three mice per group) or 8 h (LSK from three individual mice per group) followed by staining with anti-EdU and DAPI. (F) Il1 and Tnfa mRNA expression by qRT-PCR in RosaCreER+ and A20+/-RosaCreER+ LSK with 10 μM IL-1β stimulation for the indicated lengths of time (n = 3 mice per group). (G) Expression of nuclear p65 and cytoplasmic pIKKα/β by immunoblot in RosaCreER+ and A20+/-RosaCreER+ LSK BM cells with and without 15 min 10 μM IL-1β treatment. *P < 0.05. Data are represented as mean ± SEM.